Multiple injections of electroporated autologous T cells expressing a chimeric antigen receptor mediate regression of human disseminated tumor.
Multiple injections of electroporated autologous T cells expressing a chimeric antigen receptor mediate regression of human disseminated tumor.
复制标题
DOI：
10.1158/0008-5472.can-10-2880
复制
发表时间：
2010-11-15
期刊：
影响因子：
11.2
通讯作者：
June CH
中科院分区：
文献类型：
--
作者：
Zhao Y;Moon E;Carpenito C;Paulos CM;Liu X;Brennan AL;Chew A;Carroll RG;Scholler J;Levine BL;Albelda SM;June CH
通过基因转移重定向T淋巴细胞抗原特异性可以为过继性免疫治疗提供大量肿瘤反应性T淋巴细胞。然而，与病毒载体生产相关的安全问题限制了表达嵌合抗原受体（CAR）的T细胞的临床应用。 T 淋巴细胞可以通过 RNA 电穿孔进行基因修饰，而无需担心整合相关的安全问题。为了建立过继免疫治疗的安全平台，我们首先优化了RNA体外转录的载体主链，以实现高水平的转基因表达。 RNA 电穿孔 T 细胞的 CAR 表达和功能可在电穿孔后一周内检测到。多次注射 RNA CAR 电穿孔 T 细胞介导 NOD/scid/γc(−/−) 小鼠大血管化侧翼间皮瘤肿瘤的消退。当预先存在的腹膜内人源性肿瘤（已在体内生长超过 50 天）通过多次注射电穿孔抗间皮素 CAR mRNA 的自体人 T 细胞进行治疗时，肿瘤也显着减少。这是第一份使用匹配的患者肿瘤和淋巴细胞的报告，证明来自癌症患者的自体 T 细胞可以被设计为在稳健的临床前模型中为播散性肿瘤提供有效的治疗。多次注射RNA工程化T细胞是一种过继细胞转移的新方法，为癌症治疗提供了灵活的平台，可以补充逆转录病毒和慢病毒工程化T细胞的使用。这种方法可能会增加被设计来表达强大激活结构域的 T 细胞的治疗指数，而无需考虑整合病毒载体的相关安全问题。
Redirecting T lymphocyte antigen specificity by gene transfer can provide large numbers of tumor reactive T lymphocytes for adoptive immunotherapy. However, safety concerns associated with viral vector production have limited clinical application of T cells expressing chimeric antigen receptors (CARs). T lymphocytes can be gene modified by RNA electroporation without integration-associated safety concerns. To establish a safe platform for adoptive immunotherapy, we first optimized the vector backbone for RNA in vitro transcription to achieve high level transgene expression. CAR expression and function of RNA-electroporated T cells could be detected up to a week post electroporation. Multiple injections of RNA CAR electroporated T cells mediated regression of large vascularized flank mesothelioma tumors in NOD/scid/γc(−/−) mice. Dramatic tumor reduction also occurred when the pre-existing intraperitoneal human-derived tumors, that had been growing in vivo for over 50 days, were treated by multiple injections of autologous human T cells electroporated with anti-mesothelin CAR mRNA. This is the first report using matched patient tumor and lymphocytes demonstrating that autologous T cells from cancer patients can be engineered to provide an effective therapy for a disseminated tumor in a robust preclinical model. Multiple injections of RNA engineered T cells are a novel approach for adoptive cell transfer, providing flexible platform for the treatment of cancer that may complement the use of retroviral and lentiviral engineered T cells. This approach may increase the therapeutic index of T cells engineered to express powerful activation domains without the associated safety concerns of integrating viral vectors.